Extracellular vesicles-coupled miRNAs from oviduct and uterus modulate signaling pathways related to lipid metabolism and bovine early embryo development.

BACKGROUND: Extracellular vesicles (EVs) present in oviductal (OF) and uterine fluid (UF) have been shown to enhance bovine embryo quality during in vitro culture by reducing lipid contents and modulating lipid metabolism-related genes (LMGs), while also influencing cell proliferation, suggesting their involvement on the regulation of different biological pathways. The regulation of signaling pathways related to cell differentiation, proliferation, and metabolism is crucial for early embryo development and can determine the success or failure of the pregnancy. Bioactive molecules within EVs in maternal reproductive fluids, such as microRNAs (miRNAs), may contribute to this regulatory process as they modulate gene expression through post-transcriptional mechanisms. RESULTS: From the 20 differentially expressed miRNAs, 19 up-regulated in UF-EVs (bta-miR-134, bta-miR-151-3p, bta-miR-155, bta-miR-188, bta-miR-181b, bta-miR-181d, bta-miR-224, bta-miR-23b-3p, bta-miR-24-3p, bta-miR-27a-3p, bta-miR-29a, bta-miR-324, bta-miR-326, bta-miR-345-3p, bta-miR-410, bta-miR-652, bta-miR-677, bta-miR-873 and bta-miR-708) and one (bta-miR-148b) in OF-EVs. These miRNAs were predicted to modulate several pathways such as Wnt, Hippo, MAPK, and lipid metabolism and degradation. Differences in miRNAs found in OF-EVs from the early luteal phase and UF-EVs from mid-luteal phase may reflect different environments to meet the changing needs of the embryo. Additionally, miRNAs may be involved, particularly in the uterus, in the regulation of embryo lipid metabolism, immune system, and implantation. This study evaluated miRNA cargo in OF-EVs from the early luteal phase and UF-EVs from the mid-luteal phase, coinciding with embryo transit within oviduct and uterus in vivo, and its possible influence on LMGs and signaling pathways crucial for early embryo development. A total of 333 miRNAs were detected, with 11 exclusive to OF, 59 to UF, and 263 were common between both groups. CONCLUSIONS: Our study suggests that miRNAs within OF- and UF-EVs could modulate bovine embryo development and quality, providing insights into the intricate maternal-embryonic communication that might be involved in modulating lipid metabolism, immune response, and implantation during early pregnancy.

Outcomes of permanent canines on the cleft side after secondary alveolar grafting using different materials in complete unilateral cleft lip and palate.

The prevalence of impaction of the permanent canine on the cleft side (PCCS) ranges from 12-35% after alveolar bone grafting (ABG). PCCSs usually develop above other permanent teeth in the alveolar process, gradually becoming vertical until they reach the occlusal plane. The type of cleft, hypodontia of lateral incisor on the cleft side, slower PCCS root development, and genetic factors are predictors of impaction and/or its ectopic eruption. To compare the behavior of PCCS in individuals with complete unilateral cleft lip and palate (UCLP) subjected to secondary alveolar grafting (SAG) with different materials. This retrospective longitudinal study analyzed 120 individuals undergoing SAG with iliac crest bone, rhBMP-2, and mandibular symphysis. The individuals were selected at a single center and equally divided into three groups. Panoramic radiographs were analyzed by the Dolphin Imaging 11.95 software to measure PCCS angulation and PCCS height from the occlusal plane at two different timepoints. No statistical significance was found between grafting materials (P=0.416). At T1, the PCCS height from the occlusal plane was greater for rhBMP-2 and mandibular symphysis compared to iliac crest bone. The lateral incisor on the cleft side was not related to success or lack of eruption of PCCS (P=0.870). Impaction rates of PCCS were similar for the materials studied. Absence of the lateral incisor on the cleft side did not prevent spontaneous eruption of PCCSs.

Early orthognathic surgery with postsurgical bone-anchored retention in unilateral cleft lip and palate: A case report.

We present a case report of early Le Fort I osteotomy with maxillary advancement retained postoperatively by Class III elastics anchored on miniplates in a growing patient with complete unilateral cleft lip and palate (UCLP). A 14-year-old boy who underwent orthognathic surgery at the pubertal growth spurt was presented. During surgery, Bollard miniplates were installed in the posterior region of the maxilla and in the anterior region of the mandible. Class III elastics anchored on miniplates were used at night (8-10 h) starting 60 days after surgery. The force of the elastics progressively increased from 100 g to 250 g. The elastics were replaced daily. The positive overjet remained stable over 15 months of postoperative follow-up. Maxillary advancement was adequately retained using Bollard miniplates and the facial profile remained stable until the end of facial growth.

Evaluation of Radiographic Outcomes of Alveolar Graft Associated with Premaxillary Osteotomy Performed with rhBMP-2.

OBJECTIVE: To evaluate the influence of filling material and timing of surgery on radiograph outcomes of alveolar grafting with premaxillary osteotomy. The null hypothesis was that radiographic outcomes would be similar with both rhBMP-2 (rhBMP-2G) and cancellous bone from the iliac crest (IG), regardless of the timing of surgery. DESIGN: Cross-sectional study with consecutive sampling of 56 periapical or occlusal radiographs taken 12 months after surgery. SETTING: A single tertiary craniofacial center. PATIENTS/PARTICIPANTS: Twenty-eight patients with complete bilateral cleft lip and palate and mean age of 13 years. The individuals underwent bilateral alveolar grafting associated with premaxillary osteotomy (AG + PO) with rhBMP-2 or cancellous bone from the iliac crest. INTERVENTIONS: Experienced maxillofacial surgeons used the same surgical technique in both groups. AG + PO were assigned as success or failure by 3 blinded raters based on modified Bergland and SWAG scales. MAIN OUTCOME MEASURES: The influence of filling materials and timing of surgery on radiographic outcomes was verified by Fisher's exact test and chi-square test (P < .05). RESULTS: There was no significance variation between the mean age of participants in the rhBMP-2G and IG (P = .471). Scales showed almost perfect reliability (agreement rate = 96.4%; K = 0.85). rhBMP-2G and IG had similar success rates with modified Bergland scale (85.7% and 82.1%) and SWAG scale (92.9% and 82.1%), respectively. However, only modified Bergland scale found influence of age on radiographic outcomes (P = .025). CONCLUSIONS: AG + PO performed with rhBMP-2 and iliac crest bone showed similar radiographic success rates, regardless of the timing of surgery.

Avaliação da satisfação com a vida e o risco para violência em idosos / Assessment of life satisfaction and risk of violence in elderly / Evaluación de la satisfacción con la vida y riesgo de violencia en personas mayores

Objetivo:descrever a relação da satisfação com a vida e o risco para violência entre homens e mulheres idosos. Método: estudo quantitativo, transversal, realizado com 159 idosos, utilizando a Escala de Satisfação com a Vida e Hwalek-Sengstock Elder Abuse Screening Test. Os dados foram analisados mediante estatística descritiva (frequência absoluta e relativa, medidas de tendência central e dispersão) e inferencial (Teste t de Student; Regressão Linear). Resultados: a satisfação com a vida segundo o gênero apresentou diferença significativa entre as médias do risco para violência, verificando que os homens idosos que apresentavam risco para violência estavam menos satisfeitos com a vida. Observou-se correlação negativa para o grupo masculino (ß=-0,416) e feminino (ß=-0,293) entre essas variáveis. Conclusão: homens idosos que apresentam risco para violência exibem menores escores de satisfação, apontando que a satisfação com a vida aumenta à medida em que o risco para violência diminui

Objective: to describe the relationship between life satisfaction and risk of violence among elderly men and women. Method: quantitative study, cross-sectional, conducted with 159 elderlies, using the Life Satisfaction Scale and Hwalek-Sengstock Elder Abuse Screening Test. Data were analyzed through descriptive statistic (absolute and relative frequency, measures of central tendency and dispersion) and inferential (Student's t-test; Linear regression). Results: life satisfaction according to gender presented significant difference between the averages of the risk of violence, verifying that older men that presented risk of violence were less satisfied with life. It was observed negative correlation for male group (ß=-0,416) and female between these variables. (ß=-0,293). Conclusion: older men that presented risk of violence exhibit lower satisfaction scores, pointing out that life satisfaction increases as the risk of violence decreases

Objetivo: describir la relación entre la satisfacción vital y el riesgo de violencia entre hombres y mujeres ancianos. Método: estudio cuantitativo, transversal, realizado con 159 ancianos, utilizando la Escala de Satisfacción con la Vida y Hwalek-Sengstock Elder Abuse Screening Test. Los datos se analizaron mediante estadísticas descriptivas (frecuencia absoluta y relativa, medidas de tendencia central y dispersión) y estadísticas inferenciales (prueba t de Student; regresión lineal). Resultados: la satisfacción con la vida según el género mostró diferencia significativa entre las medias de riesgo de violencia, verificando que los hombres ancianos que presentaban riesgo de violencia estaban menos satisfechos con la vida. Se observó correlación negativa para el grupo de hombres (ß=-0,416) y mujeres (ß=-0,293) entre estas variables. Conclusión: los hombres ancianos que presentan riesgo de violencia muestran puntuaciones de satisfacción más bajas, lo que indica que la satisfacción con la vida aumenta a medida que disminuye el riesgo de violencia

Outcomes of permanent canines on the cleft side after secondary alveolar grafting using different materials in complete unilateral cleft lip and palate

Abstract The prevalence of impaction of the permanent canine on the cleft side (PCCS) ranges from 12-35% after alveolar bone grafting (ABG). PCCSs usually develop above other permanent teeth in the alveolar process, gradually becoming vertical until they reach the occlusal plane. The type of cleft, hypodontia of lateral incisor on the cleft side, slower PCCS root development, and genetic factors are predictors of impaction and/or its ectopic eruption. Objective: To compare the behavior of PCCS in individuals with complete unilateral cleft lip and palate (UCLP) subjected to secondary alveolar grafting (SAG) with different materials. Methodology: This retrospective longitudinal study analyzed 120 individuals undergoing SAG with iliac crest bone, rhBMP-2, and mandibular symphysis. The individuals were selected at a single center and equally divided into three groups. Panoramic radiographs were analyzed by the Dolphin Imaging 11.95 software to measure PCCS angulation and PCCS height from the occlusal plane at two different timepoints. Results: No statistical significance was found between grafting materials (P=0.416). At T1, the PCCS height from the occlusal plane was greater for rhBMP-2 and mandibular symphysis compared to iliac crest bone. The lateral incisor on the cleft side was not related to success or lack of eruption of PCCS (P=0.870). Conclusion: Impaction rates of PCCS were similar for the materials studied. Absence of the lateral incisor on the cleft side did not prevent spontaneous eruption of PCCSs.

Novel Combination of the Biophysical, Nutritional, and Nutraceutical Properties in Subtropical Pigmented Maize Hybrids.

Maize (Zea mays L.) represents the main caloric source for much of the world's population. Pigmented maize varieties are an excellent source of nutraceutical compounds: blue and yellow maize are rich in anthocyanins as well as carotenoids and phenolic acids, respectively. However, blue maize is usually grown in small quantities as a specialty crop because it lacks the qualities and adaptations of commercial white and yellow varieties. Here, a new high-yield variety of blue maize called Vitamaiz was developed from inbred lines of subtropical blue, white, and yellow maize. The aim of this study was to characterize the nutraceutical and physical properties of 30 Vitamaiz hybrids in two subtropical locations. Kernel physical traits, nutrient composition, and nutraceutical components (free phenolic acids, FPA; cell wall-bound phenolic acids, BPA; total monomeric anthocyanin content, TAC; antioxidant capacity, AOX by oxygen radical absorbance capacity assay, and total carotenoid content, TCC) were evaluated. The biophysical traits of the hybrids were suitable for nixtamalized and flour maize industries. High levels of FPA (228 mg GAE/100 g), BPA (635 mg GAE/100 g), and AOX (2.0 and 8.1 mM Trolox equivalent/100 g for FPA and BPA, respectively) were also detected with elevated TAC levels (274 mg C3G/kg dw) and AOX activity (3.1 mM Trolox equivalent/100 g). This is the first study to characterize Blue × Yellow maize hybrids that adapt to subtropical environments.

Extracellular vesicles from oviductal and uterine fluids supplementation in sequential in vitro culture improves bovine embryo quality.

BACKGROUND: In vitro production of bovine embryos is a well-established technology, but the in vitro culture (IVC) system still warrants improvements, especially regarding embryo quality. This study aimed to evaluate the effect of extracellular vesicles (EVs) isolated from oviductal (OF) and uterine fluid (UF) in sequential IVC on the development and quality of bovine embryos. Zygotes were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum (dFCS) in the presence (BSA-EV and dFCS-EV) or absence of EVs from OF (D1 to D4) and UF (D5 to D8), mimicking in vivo conditions. EVs from oviducts (early luteal phase) and uterine horns (mid-luteal phase) from slaughtered heifers were isolated by size exclusion chromatography. Blastocyst rate was recorded on days 7-8 and their quality was assessed based on lipid contents, mitochondrial activity and total cell numbers, as well as survival rate after vitrification. Relative mRNA abundance for lipid metabolism-related transcripts and levels of phosphorylated hormone-sensitive lipase (pHSL) proteins were also determined. Additionally, the expression levels of 383 miRNA in OF- and UF-EVs were assessed by qRT-PCR. RESULTS: Blastocyst yield was lower (P < 0.05) in BSA treatments compared with dFCS treatments. Survival rates after vitrification/warming were improved in dFCS-EVs (P < 0.05). EVs increased (P < 0.05) blastocysts total cell number in dFCS-EV and BSA-EV compared with respective controls (dFCS and BSA), while lipid content was decreased in dFCS-EV (P < 0.05) and mitochondrial activity did not change (P > 0.05). Lipid metabolism transcripts were affected by EVs and showed interaction with type of protein source in medium (PPARGC1B, LDLR, CD36, FASN and PNPLA2, P < 0.05). Levels of pHSL were lower in dFCS (P < 0.05). Twenty miRNA were differentially expressed between OF- and UF-EVs and only bta-miR-148b was increased in OF-EVs (P < 0.05). CONCLUSIONS: Mimicking physiological conditions using EVs from OF and UF in sequential IVC does not affect embryo development but improves blastocyst quality regarding survival rate after vitrification/warming, total cell number, lipid content, and relative changes in expression of lipid metabolism transcripts and lipase activation. Finally, EVs miRNA contents may contribute to the observed effects.

XPC-PARP complexes engage the chromatin remodeler ALC1 to catalyze global genome DNA damage repair.

Cells employ global genome nucleotide excision repair (GGR) to eliminate a broad spectrum of DNA lesions, including those induced by UV light. The lesion-recognition factor XPC initiates repair of helix-destabilizing DNA lesions, but binds poorly to lesions such as CPDs that do not destabilize DNA. How difficult-to-repair lesions are detected in chromatin is unknown. Here, we identify the poly-(ADP-ribose) polymerases PARP1 and PARP2 as constitutive interactors of XPC. Their interaction results in the XPC-stimulated synthesis of poly-(ADP-ribose) (PAR) by PARP1 at UV lesions, which in turn enables the recruitment and activation of the PAR-regulated chromatin remodeler ALC1. PARP2, on the other hand, modulates the retention of ALC1 at DNA damage sites. Notably, ALC1 mediates chromatin expansion at UV-induced DNA lesions, leading to the timely clearing of CPD lesions. Thus, we reveal how chromatin containing difficult-to-repair DNA lesions is primed for repair, providing insight into mechanisms of chromatin plasticity during GGR.

Application of metabolic engineering to enhance the content of alkaloids in medicinal plants.

Plants are a rich source of bioactive compounds, many of which have been exploited for cosmetic, nutritional, and medicinal purposes. Through the characterization of metabolic pathways, as well as the mechanisms responsible for the accumulation of secondary metabolites, researchers have been able to increase the production of bioactive compounds in different plant species for research and commercial applications. The intent of the current review is to describe the metabolic engineering methods that have been used to transform in vitro or field-grown medicinal plants over the last decade and to identify the most effective approaches to increase the production of alkaloids. The articles summarized were categorized into six groups: endogenous enzyme overexpression, foreign enzyme overexpression, transcription factor overexpression, gene silencing, genome editing, and co-overexpression. We conclude that, because of the complex and multi-step nature of biosynthetic pathways, the approach that has been most commonly used to increase the biosynthesis of alkaloids, and the most effective in terms of fold increase, is the co-overexpression of two or more rate-limiting enzymes followed by the manipulation of regulatory genes.

Case Report: Depression × dementia with Lewy bodies in the elderly: The importance of differential diagnosis.

Background: Dementia is a clinical syndrome which is more common in elderly people. Dementia with Lewy bodies (LBD) is not so rare in elderly people, with cognitive impairment in about 30% over age 65. The clinical picture is characterized by fluctuation in cognitive functions, recurrent, well-formed, detailed visual hallucinations, and Parkinsonism, with rigidity, tremor, bradykinesia, and slurred speech. Case presentation: We present a case report of LBD in a 73-year-old retired teacher, which a initial wrong diagnosis of refractory depression for at least 3 years. We also conduct a review of recent works on theme. Conclusion: LBD diagnosis can be neglected for years, with a legal and clinical issues to patients and their families. Detailed medical research, including differential diagnosis, are very necessary on those cases, specially when they are called refractory. We encourage new research and adequate clinical training to prevent damage.

Effects of different stimulators of cGMP synthesis on lipid content in bovine oocytes matured in vitro.

Bovine oocytes and blastocysts produced in vitro are frequently of lower quality and less cryotolerant than those produced in vivo, and greater accumulation of lipids in the cytoplasm has been pointed out as one of the reasons. In human adipocytes cGMP signaling through the activation of PKG appears to be involved in lipid metabolism, and components of this pathway have been detected in bovine cumulus-oocyte complexes (COCs). The aim of this study was to investigate the influence of this pathway on the lipid content in oocytes and expression of PLIN2 (a lipid metabolism-related gene) in cumulus cells. COCs were matured in vitro for 24 h with different stimulators of cGMP synthesis. The activation of soluble guanylyl cyclase (sGC) by Protoporphyrin IX reduced lipid content (22.7 FI) compared to control oocytes (36.45 FI; P <0.05). Stimulation of membrane guanylyl cyclase (mGC) with natriuretic peptides precursors A and C (NPPA and NPPC) had no effect (36.5 FI; P>0.05). When the PKG inhibitor KT5823 was associated with Protoporphyrin IX, its effect was reversed and lipid contents increased (52.71 FI; P<0.05). None of the stimulators of cGMP synthesis affected the expression of PLIN2 in cumulus cells. In conclusion, stimulation of sGC for cGMP synthesis promotes lipolytic activities in bovine oocytes matured in vitro and such effect is mediated by PKG. However, such effect may vary depending on the stimulus received and/or which synthesis enzyme was activated, as stimulation of mGC had no effects.

Wild and trapped: a history of Colombian zoos and its revelations of animal fortunes and State entanglements, 1930s-1990s / Selvagens e confinados: uma história dos zoológicos colombianos e suas revelações sobre os destinos de animais e entrelaçamentos do Estado, décadas de 1930 a 1990

Abstract This article presents the first history of Colombian zoos and one of the few contributions to the history of these institutions in Latin America. It proposes that in this region's zoos native animals tended to predominate, signaling the increasing transformation of forests and other native ecosystems into croplands and pasturelands, as well as the growing distance between people and natural environments. Colombian zoos additionally underscore the limitations of the State in its period of most rapid expansion. They reveal how private interests overshadowed the State by providing public amenities, but also how they offered unconventional solutions to certain hurdles of State formation.

Resumo Este artigo apresenta a primeira história dos zoológicos colombianos e é uma das poucas contribuições à história dessas instituições na América Latina. Nele, propõe-se que nos zoológicos dessa região predominavam animais nativos, indício da crescente transformação de florestas e outros ecossistemas nativos em terrenos agrícolas e pastos, assim como do gradativo distanciamento entre pessoas e natureza. Os zoológicos colombianos também evidenciam as limitações do Estado no período de sua mais rápida expansão. Eles revelam como os interesses privados eclipsaram o Estado fornecendo comodidades públicas, mas também como ofereceram soluções não convencionais a algumas das dificuldades na formação do Estado.

Characterization and profiling analysis of bovine oviduct and uterine extracellular vesicles and their miRNA cargo through the estrous cycle.

Extracellular vesicles (EVs) found in various biological fluids and particularly in reproductive fluids, have gained considerable attention for their possible role in cell- to- cell communication. Among, the different bioactive molecules cargos of EVs, MicroRNAs (miRNAs) are emerging as promising diagnostic biomarkers with high clinical potential. Aiming to understand the roles of EVs in bovine reproductive tract, we intended to characterize and profile the EVs of oviduct and uterine fluids (OF-EVs, UF-EVs) and their miRNA across the estrous cycle. Nanoparticle tracking analysis and transmission electron microscopy confirmed the existence of small EV population in OF and UF at all stages, (size between 30 and 200 nm; concentration: 3.4 × 1010  EVs/ml and 6.0 × 1010  EVs/ml for OF and UF, respectively, regardless of stage). The identification of EV markers (CD9, HSP70, and ALIX proteins) was confirmed by western blot. The miRNA analysis revealed the abundance of 310 and 351 miRNAs in OF-EVs and UF-EVs, respectively. Nine miRNAs were differentially abundant in OF-EVs between stages of the cycle, eight of them displayed a progressive increase from S1 to S4 (p < .05). In UF-EVs, a total of 14 miRNAs were differentially abundant between stages. Greater differences were observed between stage 1 (S1) and stage 3 (S3), with 11 miRNAs enriched in S3 compared to S1. Functional enrichment analysis revealed the involvement of these miRNAs in relevant pathways such as cell signaling, intercellular junctions, and reproductive functions that may be implicated in oviduct and uterus modulation across the cycle, but also in their preparation for embryo/conceptus presence and development.

Nobiletin enhances the development and quality of bovine embryos in vitro during two key periods of embryonic genome activation.

In vitro culture can alter the development and quality of bovine embryos. Therefore, we aimed to evaluate whether nobiletin supplementation during EGA improves embryonic development and blastocyst quality and if it affects PI3K/AKT signaling pathway. In vitro zygotes were cultured in SOF + 5% FCS (Control) or supplemented with 5, 10 or 25 µM nobiletin (Nob5, Nob10, Nob25) or with 0.03% dimethyl-sulfoxide (CDMSO) during minor (2 to 8-cell stage; MNEGA) or major (8 to 16-cell stage; MJEGA) EGA phase. Blastocyst yield on Day 8 was higher in Nob5 (42.7 ± 1.0%) and Nob10 (44.4 ± 1.3%) for MNEGA phase and in Nob10 (61.0 ± 0.8%) for MJEGA phase compared to other groups. Mitochondrial activity was higher and lipid content was reduced in blastocysts produced with nobiletin, irrespective of EGA phase. The mRNA abundance of CDK2, H3-3B, H3-3A, GPX1, NFE2L2 and PPARα transcripts was increased in 8-cells, 16-cells and blastocysts from nobiletin groups. Immunofluorescence analysis revealed immunoreactive proteins for p-AKT forms (Thr308 and Ser473) in bovine blastocysts produced with nobiletin. In conclusion, nobiletin supplementation during EGA has a positive effect on preimplantation bovine embryonic development in vitro and corroborates on the quality improvement of the produced blastocysts which could be modulated by the activation of AKT signaling pathway.

Interventions to prevent elder abuse in the community: a mixed-methods systematic review.

OBJECTIVE: To identify the interventions proposed in the literature aimed at preventing elder abuse among community-residing elders. METHOD: This was a mixed-methods systematic review that followed the Preferred Reporting Items for Systematic Review and Meta-analysis Protocols checklist. The protocol of this systematic review was registered in the International Prospective Register of Systematic Reviews. We included studies published between January 2004 and December 2019 in the databases: LILACS, IBECS, CUMED, CINAHL, MEDLINE and SciELO. The methodological quality of each study included was performed using the Mixed Methods Appraisal Tool. RESULTS: Seven studies formed the final sample. Two groups emerged after the analyses: the primary level intervention group, which used health education and coexisting elder abuse groups; and the secondary intervention level, focused mostly on the mental health care of victims of elder abuse. CONCLUSION: There is still a shortage of intervention studies to prevent elder abuse. All the studies selected had a positive outcome, and all interventions can be implemented in nursing care practice.

A triskelion of nucleic acids drives protein aggregation in A-T.

Mutations in ataxia telangiectasia mutated (ATM) kinase lead to cerebellar neurodegeneration. In this issue of Molecular Cell, Lee et al. (2021) revealed how transcription-induced reactive oxygen species and DNA-RNA hybrids activate PARP enzymes, generating the nucleic acid poly-ADP-ribose, which promotes the accumulation of protein aggregates in A-T-like disorders.

Effects of saline elicitors on saponin production in Agave salmiana plants grown in vitro.

Agave plants are natives of Mexico and have an important role in the functional food industry. Agave salmiana grows in dry and desert soils, which are high in salt content; however, little is known about its response to saline conditions. In this study, A. salmiana plants grown in vitro were exposed to 0.1, 0.5, and 1.0 mM of salt elicitors, including AlCl3, NaCl, and CoCl2, and saponin synthesis and morphological characteristics were examined. Saponins were identified and quantified in ethanolic extracts using HPLC-ELSD. Root length and number, leaf length and number, and plant fresh weight were evaluated to determine the phenological condition of the plant. The presence of salts at various concentrations did not affect the physiological characteristics of the plant. Moreover, 0.5 mM NaCl induced a higher production of total saponin. Chlorogenin glycoside 1 (CG1) and hecogenin glycoside 1 (HG1) content remained unchanged across treatments. By contrast, CG2 and HG2 concentrations tended to decrease in response to increased concentrations of AlCl3, NaCl, or CoCl2. In vitro salt elicitors could be a feasible tool in the synthesis of specific saponins, without compromising on plant biomass. Our findings can be used in further generation of low saponin agave plants in field for the improvement of fermentation yield.

Isolation, Characterization, and MicroRNA Analysis of Extracellular Vesicles from Bovine Oviduct and Uterine Fluids.

Intercellular communication can be carried out by circulating systemic and/or locally released extracellular vesicles (EVs), produced by nearly every cell type and tissue, and are involved in physiological and pathological processes. In recent years, EVs have been identified in reproductive tissues, such as oviduct and uterus, and have been shown to be related to several events important for reproductive success. The understanding of their functions in reproduction has important implications for assisted reproductive technologies, for the treatment of infertility in humans and improvement of reproduction efficiency in animals. To study such EVs, it is necessary to isolate and concentrate them from fluid samples, which in the case of reproductive tissues, are usually of limited volume. Several methods for EV isolation are available such as chromatography, ultracentrifugation, polymer-based precipitation, and immunoaffinity.Outcomes can be variable in terms of the amount and quality of isolated EVs, due to the type of isolation method. The choice of method, or a different combination of methods, may depend on the type of sample and scientific question to be addressed in a given study. In this chapter, we describe a method for isolation of EVs from bovine oviductal and uterine fluids for use in functional studies. The method combines size exclusion chromatography and ultracentrifugation. We also describe the different protocols for characterization of isolated EVs (transmission electron microscopy, nanoparticle tracking analysis, and western blot), as well as the isolation of RNA content in EVs, and their miRNAs profiling for functional studies.